Development of multiplex PCR for simultaneous detection of six swine DNA and RNA viruses

Journal of Virological Methods
Xingang XuHung-Jen Liu

Abstract

Uniplex and multiplex reverse transcription-polymerase chain reaction (RT-PCR) and PCR protocols were developed and evaluated subsequently for its effectiveness in detecting simultaneously single and mixed infections in swine. Specific primers for three DNA viruses and three RNA viruses, including classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), Japanese encephalitis virus (JEV), porcine circovirus type 2 (PCV2), porcine pseudorabies virus (PRV) and porcine parvovirus (PPV) were used for testing procedure. A single nucleic acid extraction protocol was adopted for the simultaneous extraction of both RNA and DNA viruses. The multiplex PCR consisted with two-step procedure which included reverse transcription of RNA virus and multiplex PCR of viral cDNA and DNA. The multiplex PCR assay was shown to be sensitive detecting at least 450pg of viral genomic DNA or RNA from a mixture of six viruses in a reaction. The assay was also highly specific in detecting one or more of the same viruses in various combinations in specimens. Thirty clinical samples and aborted fetuses collected from 4- to 12-week-old piglets were detected among 39 samples tested by both uniplex and multiplex PCR, showi...Continue Reading

References

Oct 12, 2000·Clinical Microbiology Reviews·E M ElnifroP E Klapper
Jan 27, 2004·Veterinary Microbiology·J EllisG M Allan
Jun 30, 2004·Veterinary Microbiology·Chienjin HuangMaw-Sheng Chien
Jul 24, 2010·Virus Genes·Jie SongBaohua Zhao

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Citations

Dec 15, 2015·Transboundary and Emerging Diseases·O LungA Ambagala
Sep 10, 2014·Journal of Virological Methods·Kerstin WernikeBernd Hoffmann
May 6, 2014·Veterinary Microbiology·Yuzi LuoHua-Ji Qiu
Jan 6, 2015·Applied Microbiology and Biotechnology·Ru ChenYong-Chang Cao
Jul 22, 2017·Journal of Virological Methods·Yongjuan WangWeiyong Zuo
Dec 21, 2017·Viruses·István MészárosZoltán Zádori
Jun 3, 2018·The Journal of Immunology : Official Journal of the American Association of Immunologists·Qian DuYong Huang
Jun 30, 2019·Frontiers in Veterinary Science·Bin Zhou
Nov 23, 2019·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Nicole B GoeckeLars E Larsen
Dec 21, 2018·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Nanhua ChenJianzhong Zhu
Mar 15, 2014·Journal of Agricultural and Food Chemistry·David DobnikMaja Ravnikar
Oct 29, 2021·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Qi ZhangXingang Xu

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